Aug 26, 2023

Public workspaceDifferentiation of SH-SY5Y cells

DOI
dx.doi.org/10.17504/protocols.io.dm6gp3jj1vzp/v1
  • 1Department of Molecular and Cell Biology, Howard Hughes Medical Institute, University of California, Berkeley, Berkeley, United States;
  • 2Helen Wills Neuroscience Institute, University of California, Berkeley, Berkeley, United States
Nancy C. Hernandez Villegas
Icon indicating open access to content
QR code linking to this content
DOI: dx.doi.org/10.17504/protocols.io.dm6gp3jj1vzp/v1
Protocol Citation: Shenjie Wu, Nancy C. Hernandez Villegas, schekman 2023. Differentiation of SH-SY5Y cells. protocols.io https://dx.doi.org/10.17504/protocols.io.dm6gp3jj1vzp/v1
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License,  which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: August 04, 2023
Last Modified: August 26, 2023
Protocol Integer ID: 85961
Funders Acknowledgement:
Nancy C Hernandez Villegas
Grant ID: NIH training program T32GM139780
Randy Schekman
Grant ID: Howard Hughes Medical Institute
Abstract
This protocol describes a standard procedure to differentiate SH-SY5Y cells into dopaminergic neurons using retinoic acid
Materials
Reagents

ReagentCatalog numberManufacturer
SH-SY5Y cellsCell Culture Facility, UC Berkeley
DMEM, High Glucose, GlutaMAX Supplement10566-024Thermo Scientific
Mem Non Essential Amino Acids Solution (100X)11140050Thermo Scientific
Corning® 100 mL Sodium Pyruvate, Liquid 100 mMMT25000CICorning
FBS
Retinoic acidR2625Sigma

Differentiation of SH-SY5Y cells
Differentiation of SH-SY5Y cells
SH-SY5Y neuroblastoma cells were maintained in DMEM supplemented with 1× nonessential amino
acid (NEAA), 1× sodium pyruvate, and 10% FBS.
Differentiation was induced by lowering the FBS in culture medium to 1% plus 10 μM RA.
Cell medium was replaced each 3 days to replenish RA.
Cell morphology was monitored by microscopy and experiments on SH-SY5Y cells were performed from D6 of differentiation.